Ssp 25

Three human ICC cell lines, HCCC-9810, RBE and SSP-25, were purchased from the Cell Bank of Chinese Academy of Sciences (Shanghai, China). The cells were cultured in RPMI-1640 medium (Gibco, USA) contained 10% fetal bovine serum (FBS) (Gibco, USA), 100 U/ml penicillin (Gibco, USA), and 100 μg/ml streptomycin (Gibco, USA) in a humidified incubator at 37°C with 5% CO₂. The short hairpin FoxM1 lentiviral vectors (GeneChem Co., Ltd. Shanghai, China) designed to downregulate the expression of FoxM1 were transfected into SSP-25 cells, which were then designated SSP-25-shFoxM1. The lentiviral vectors with FoxM1 were transfected into HCCC-9810 cells, which were then designated HCCC-9810-FoxM1. Meanwhile, the empty lentiviral vectors were transfected into SSP-25 and HCCC-9810 cells as controls, which were designated SSP-25-control and HCCC-9810-control, respectively. The lentiviral vectors carried puromycin resistance and green fluorescent sequences. Subsequently, puromycin (1 µg/ml) was used for screening the stable cell lines. The transfection procedure was performed based strictly on the manufacturer’s instructions. The short hairpin RNA (shRNA) and cDNA clone sequences are listed in Supplementary Table S3.